title: In vitro strain adaptation of CWD prions by serial protein misfolding cyclic amplification.
abstract: We used serial protein misfolding cyclic amplification (sPMCA) to amplify the D10 strain of CWD prions in a linear relationship over two logs of D10 dilutions. The resultant PMCA-amplified D10 induced terminal TSE disease in CWD-susceptible Tg(cerPrP)1536 mice with a survival time approximately 80 days shorter than the original D10 inoculum, similar to that produced by in vivo sub-passage of D10 in Tg(cerPrP)1536 mice. Both in vitro-amplified and mouse-passaged D10 produced brain lesion profiles, glycoform ratios and conformational stabilities significantly different than those produced by the original D10 inoculum in Tg(cerPrP)1536 mice. These findings demonstrate that sPMCA can amplify and adapt prion strains in vitro as effectively and much more quickly than in vivo strain adaptation by mouse passage. Thus sPMCA may represent a powerful tool to assess prion strain adaptation and species barriers in vitro.
authors.affiliation: Department of Microbiology, Immunology, and Pathology, Colorado State University, College of Veterinary Medicine and Biomedical Sciences, Fort Collins, CO 80523, USA.
authors.forename: Crystal
authors.initials: C
authors.lastname: Meyerett
authors.forename: Brady
authors.initials: B
authors.lastname: Michel
authors.forename: Bruce
authors.initials: B
authors.lastname: Pulford
authors.forename: Terry R
authors.initials: TR
authors.lastname: Spraker
authors.forename: Traci A
authors.initials: TA
authors.lastname: Nichols
authors.forename: Theodore
authors.initials: T
authors.lastname: Johnson
authors.forename: Timothy
authors.initials: T
authors.lastname: Kurt
authors.forename: Edward A
authors.initials: EA
authors.lastname: Hoover
authors.forename: Glenn C
authors.initials: GC
authors.lastname: Telling
authors.forename: Mark D
authors.initials: MD
authors.lastname: Zabel
mesh_terms: D000222:Adaptation, Physiological; D000818:Animals; D001921:Brain / Q000473:pathology; D051379:Mice; D008822:Mice, Transgenic; D018620:PrPSc Proteins / Q000737:chemistry / Q000302:isolation & purification; D011328:Prions / Q000737:chemistry* / Q000302:isolation & purification / Q000502:physiology; D011487:Protein Conformation; D017510:Protein Folding; D034081:Wasting Disease, Chronic / Q000209:etiology / Q000235:genetics* / Q000473:pathology / Q000503:physiopathology*
chemical_list: D018620:PrPSc Proteins; D011328:Prions